Methods for isolation of endothelial and smooth muscle cells and in vitro proliferation assays

Angiogenesis, the formation of new blood vessel from pre-existing blood vessel, occurs in a variety of normal and pathological conditions. It is complex morphogenetic process involving the coordinate migration, invasion, and reorganization of several cell types including endothelial cells, pericytes, smooth muscle cells, and stromal fibroblasts. The angiogenic response begins with excess protease secretion to facilitate basement membrane remodeling, proliferation of endothelial cells, and endothelial cell migration to form capillary network and lumen closure. In this chapter, we describe the methods to isolate mouse and human endothelial cells, smooth muscle cells, which will provide abundant, convenient and useful tool for the investigation of many aspects of endothelial cell biology. The high degrees of functional diversity have been observed from endothelial cells derived from different organs, and within the different vascular beds of a given organ. Therefore, this apparent heterogeneity has highlighted the requirement for the endothelial cell isolation and culture from a variety of tissues of different species in order to establish more realistic in vitro angiogenic models.